Method and composition for the treatment of diarrhea and gastrointestinal spasms

ABSTRACT

Anti-diarrheal and/or gastrointestinal anti-spasmodic pharmaceutical compositions containing [A] a polyamine of the formula: 
     R 1 —N 1 (R 2 )—(CH 2 )x-N 2 H—Q—N 3 H—(CH 2 )y-N 4 (R 3 )—R 4   (I) 
     wherein: R 1 , R 2 , R 3  and R 4  may be the same or different and are H, alkyl, cycloalkyl or aralkyl having from 1 to 12 carbon atoms, or a heterocyclic group having from 3 to 10 atoms wherein the hetero atom is said N 1  or N 4 ;  
     Q is a cycloalkyl group having from 3 to 10 carbon atoms;  
     x is an integer from 3 to 6, inclusive;  
     and y is an integer from 3 to 6, inclusive;  
     or (II) a salt thereof with a pharmaceutically acceptable acid; and [B] a pharmaceutically acceptable carrier therefor as well as methods of treatment utilizing the polyamines are disclosed.

[0001] This invention was made with United States Government supportunder Grant NCDDG-CA37606, awarded by the National Cancer Institute. TheUnited States Government has certain rights in this invention.

BACKGROUND OF THE INVENTION

[0002] The present invention relates to certain novel anti-diarrheal andgastrointestinal anti-spasmodic agents and methods of treatment andpharmaceutical compositions based thereon.

DESCRIPTION OF THE PRIOR ART

[0003] Diarrhea can result from a variety of pathophysiologicaldisorders including bacterial and parasitic infections, disease ordebilitation of organs such as liver, adrenal and others. It can alsooccur as a result of other therapy or diet. In all cases, diarrhea isgenerally a symptom of organic gastrointestinal disorders and not itselfa disorder. Chronic diarrhea is generally due to: (1) hypersecretion offluid and electrolytes of the stomach, small intestine and colon; (2)inability to absorb certain nutrients (malabsorption); and (3)intestinal hypermotility and rapid transport. These may occur separatelyor in combination. Certain disorders may have diarrhea as a prominentfeature of the disease/syndrome, but the specific etiology is unclear.In this latter group, emotional tension and psychological factors mayadversely influence the frequency of the symptoms.

[0004] Diarrhea and diarrheal diseases are one of the most frequentcauses of morbidity and mortality, especially in less developedcountries wherein the number of those killed by such diseases isestimated at about 5 million persons per annum. Particularly dangerousare diarrheal diseases of the newborn and the youngest group of babies(S. Hughes: Drugs, Vol. 26, pp. 80-90 (1983)).

[0005] In mechanized or automated large capacity farms, diarrhea andinfections of the gastrointestinal tract are frequent, especially withyoung livestock and the high mortality or growth deceleration thereofhave a considerable negative economical effect. Diarrheal diseases ofman and animals are caused by a plurality of etiological factors,especially of microbial and viral character. The most prevalent microbesare gram-negative bacteria, Escherichia coli and Vibrio cholerae.However, it is now clear that other bacteria, viruses and parasites(protozoan, amoeba, etc.) also cause severe problems.

[0006] Diarrheal diseases are treated by rehydration therapy usingpreparations composed of various salts (potassium chloride, sodiumchloride, sodium hydrogen carbonate) and glucose, whereby quickcompensation for the loss of water and ions, as well as for acidosis,occurs. However, the occurrence of diarrheal diseases is not influenced.Other substances of the same kind produce similar results.

[0007] Anti-diarrheal compounds are, of course, well known in themedicinal arts and take various forms. In particular, there are avariety of products known which act systemically to provideanti-diarrheal effects when administered in a manner which will enablethe drug to be taken into the system at effective therapeutic levels.

[0008] In addition, there are anti-cholinergic substances appliedtogether with spasmolytics such as Reasec. RTM. (Janssen) which containdiphenyloxylate and atropin. Both human and veterinary medicine usechemotherapeutic agents with anti-bacterial effects, such assulfonamides, or antibiotics are availed of which are apt to suppresscertain infections.

[0009] Medicaments are also aimed at the sphere of regulation dependingon receptors, especially those localized on the basolateral membrane,further by means of an intracellular mechanism of intervention by theso-called secondary messenger, and by influencing the transportmechanism, especially boundary membranes. The modulation ofreceptor-dependent regulation mechanisms can be influenced, to someextent, by medicaments of the type alpha.sub.2 adrenergic agonists suchas clonidine (Catapresan.RTM.) (E. B. Chang et al, Gastroenterology,Vol. 91, pp. 564-569 (1986)), somatostatin, or encephalin and morphineanalogs. For influencing the transport of ions through the membrane, itis also possible to use alpha.sub.2 adrenergic agonists (E. B. Chang etal, Am. J. Physiol., Vol. 1982, p. 242). Reference has also been made tothe use of lidamidine, i.e., the medicament having a damping effect onthe intestine peristaltics (M. D. Dharmsathphom: Gastroenterology, Vol.91, pp. 769-775 (1986)).

[0010] Disadvantages of anti-diarrheal medicaments, i.e., those referredto in professional papers rather than those medicaments of this typeapplied in practice, include their secondary strong effects such asantihypertensive effects (clonidine), growth factors (somatostatin),habituation and/or incomplete preclinical research (encephalinderivatives). The application of large doses of antibiotics and longadministration thereof has not proved optimum in epidemical diarrhealocalities. Where the diarrhea-inducing agent is cholera toxin, however,there does not exist any efficient protection, except for inoculum whichis not sufficiently patent either, and gives short-term protection only(3 months) and low efficiency (30-40%).

[0011] In recent years, a great deal of attention has been focussed onthe polyamines, e.g., spermidine, norspermidine, homospermidine,1,4-diaminobutane (putrescine) and spermine. These studies have beenlargely directed at the biological properties of the polyamines probablybecause of the role they play in proliferative processes. It was shownearly on that the polyamine levels in dividing cells, e.g., cancercells, are much higher than in resting cells. See Janne et al, A.Biochim. Biophys. Acta., Vol. 473, p. 241 (1978); Fillingame et al,Proc. Natl. Acad. Sci. U.S.A., Vol. 72, p. 4042 (1975); Metcalf et al,J. Am. Chem. Soc., Vol. 100, p. 2551 (1978); Flink et al, Nature(London), Vol. 253, p. 62 (1975); and Pegg et al, Polyamine Metabolismand Function, Am. J. Cell. Physiol., Vol. 243, pp. 212-221 (1982).

[0012] Several lines of evidence indicate that polyamines, particularlyspermidine, are required for cell proliferation: (i) they are found ingreater amounts in growing than in non-growing tissues; (ii) prokaryoticand eukaryotic mutants deficient in polyamine biosynthesis areauxotrophic for polyamines; and (iii) inhibitors specific for polyaminebiosynthesis also inhibit cell growth. Despite this evidence, theprecise biological role of polyamines in cell proliferation isuncertain. It has been suggested that polyamines, by virtue of theircharged nature under physiological conditions and their conformationalflexibility, might serve to stabilize macromolecules such as nucleicacids by anion neutralization. See Dkystra et al, Science, Vol. 149, p.48 (1965); Russell et al, Polyamines as Biochemical Markers of Normaland Malignant Growth (Raven, New York, 1978); Hirschfield et al, J.Bacteriol., Vol. 101, p. 725 (1970); Hafner et al, J. Biol. Chem., Vol.254, p. 12419 (1979); Cohn et al, J. Bacteriol., Vol. 134, p. 208(1978); Pohjatipelto et al, Nature (London), Vol. 293, p. 475 (1981);Mamont et al, Biochem. Biophys. Res. Commun., Vol. 81, p. 58 (1978);Bloomfield et al, Polyamines in Biology and Medicine (D. R. Morris andL. J. Morton, eds., Dekker, New York, 1981), pp. 183-205; Gosule et al,Nature, Vol. 259, p. 333 (1976); Gabbay et al, Ann. N. Y. Acad. Sci.,Vol. 171, p. 810 (1970); Suwalsky et al, J. Mol. Biol., Vol. 42, p. 363(1969); and Liquori et al, J. Mol. Biol., Vol. 24, p. 113 (1968).

[0013] However, regardless of the reason for increased polyamine levels,the phenomenon can be and has been exploited in chemotherapy. SeeSjoerdsma et al, Butterworths Int. Med. Rev.: Clin. Pharmacol. Thera.,Vol. 35, p. 287 (1984); Israel et al, J. Med. Chem., Vol. 16, p. 1(1973); Morris et al, Polyamines in Biology and Medicine; Dekker, NewYork, p. 223 (198 1); and Wang et al, Biochem. Biophys. Res. Commun.,Vol. 94, p. 85 (1980).

[0014] It has been previously reported that diethylhomospermine (DEHSPM)inhibited myoelectric activity and transit of the small intestine inrats [J. Gastro. Motil., Vol. 1, p. 53 (1989)]. This inhibition wasreversed with co-administration of bethanechol, a cholinergic agonist,but not with other agonists or antagonists [Gastro., Vol. 98, p. A388(1990)]. However, there is no suggestion or disclosure in the prior artthat any of the above-described polyamines have utility asanti-diarrheal or gastrointestinal anti-spasmodic agents.

[0015] Recently, it has been found that certain polyamines functioneffectively as anti-diarrheal and gastrointestinal anti-spasmodicagents. See U.S. Pat. Nos. 5,393,757 and 5,462,970, the entire contentsand disclosures of which are incorporated herein by reference. See alsoBergeron et al, J. Med. Chem, Vol. 39, pp 2461-2471 (1996), whichdiscloses that certain hydroxy-substituted polyamines are effectiveanti-diarrheal agents. In a series of studies, Tansy was able todemonstrate that polyamiries have a profound impact on the motility ofthe gastrointestinal (UI) tract. The original work focused onpoly(etbylenimine) and gastric emptying in rodents and dogs.Branched-chain poly-(ethylenimine)s effected significant inhibition ofgastric emptying in rodents; however, they caused a severe retchresponse in dogs. Because of the structural relationship between thepoly(ethylenimine)s and natural polyamines, Tansy elected to evaluatethe effect of spermidine, spermine, and a group of polyamine analogueson the gastric emptying of rodents. It soon became clear that polyamineshad a substantial influence on gastric emptying and that “endogenousspermine and spermidine may have some unrecognized GI secretomotoractivity”. [See Spermine and Spermidine as Inhibitors ofGastrointestinal Motor Activity, Surg. Gyn. Obst, 1982, 154, 74-80;Pharmacology of Polyethylenimine I: Effects on Gastric Emptying In Rats,J. Pharm. Sci. 1977, 66. 899-901; GI Pharmacology of PolyetbyleneimineII: Motor Activity in Anesthetized Dogs, J. Pharm Sci. 1977, 66,902-904; Effects of Spermine end Spermadine on Gastric Emptying in Rats,J. Pharm. Sci. 1981, 70 347]. From a structure—activity perspective, italso became obvious that minor changes in the polyarnine's structurecould completely eradicate the molecule's ability to inhibit gastricemptying. These studies strongly suggested that the polyarninepharmacophore was an excellent candidate for the construction ofantitransit, antidiarrheal drugs.

[0016] One polyamine analogue designed and synthesized by the presentinventor, N¹, N¹⁴ diethylhomospermine (DEHSPM), is a very potentantidiarrheal. This has been demonstrated in a number of animal modelsand in the clinic against AIDS-related diarrhea. However, DEHDSPM isN-deethylated to homospermine (HSPM), which has a very protractedhalf-life: 2-3 weeks in mice and even longer in the dog. Each succeedingdose of DEHSPM results in a further accumulation of HSPM until toxiclevels of the latter tetraamine are reached, a troublesome metabolicproperty in animals. [See Structural Specificity of Synthetic Analoguesof Polyamines and their Effect on Gastrointestinal Motility, Polyaminesand the Gastrointetinal Tract, Falk Symposium, NO. 62; Kluwer Academic:Boston, 1991; Potent Anti-Diarrheal Activity of a New Class ofCompounds: Synthetic Analogues of the Polyamine Pathway.Gastroenterology 1993, 104, A54; J. Metabolism and Pharmacokinetics ofN¹, N¹⁴-Diethylhomospermine. Drug Metab. Dispos. 1996, 24, 334-843].

[0017] It is an object of the present invention to provide novelanti-diarrheal and gastrointestinal antispasmodic pharmaceuticalcompositions and methods of treatment based upon certain polyaminesother than those described in the prior art as effective therefor.

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The FIGURE is a reaction scheme for preparing the polyaminesemployed in the practice of the invention.

[0019] SYNTHESIS OF CHX(3,4,3)-trans

[0020] N,N′-Bis[3-(ethylamino)propyl]-trans-1,4-cyclohexanediaminetetrahydrochloride [CHX(3,4,3)-trans, 1] was assembled via fragmentsynthesis (Scheme 1). Disulfonamide 3, available in 90% yield fromreaction of trans-1,4-diaminocyclohexane (2) with mesitylenesulfonylchloride (2 equiv) under biphasic conditions, was alkylated withN-(3-bromopropyl)-N-ethylmesitylenesulfonamide (2 equiv, NaH, DMF) toprovide protected polyamine 4 in 73% yield. The amino groups of 4 wereunmasked with 30% HBr in acetic acid and phenol, furnishing finalproduct 1 in 64% yield. [Bergeron et al, J. Med. Chem., Vol. 7,pp3464-3476 (1984)]

SUMMARY OF THE INVENTION

[0021] The foregoing and other objects are realized by the presentinvention, one embodiment of which is an anti-diarrheal, anti-secretory,or gastrointestinal anti-spasmodic pharmaceutical composition comprisingan anti-diarrheal or gastrointestinal antispasmodic (hereinafter “GIanti-spasmodic”) effective amount of a compound of the formula set forthbelow and a pharmaceutically acceptable carrier therefor.

[0022] An additional embodiment of the present invention comprises amethod of treating a human or nonhuman animal in need thereof comprisingadministering to the animal an anti-diarrheal or GI antispasmodiceffective amount of a compound of the formulae below.

[0023] Suitable methods for the preparation of polyamines for use in thecomposition and method of the invention are those described inapplication Ser. No. 07/210,520 filed Jun. 23, 1988, now U.S. Pat. No.5,091,576, the entire contents and disclosure of which is incorporatedherein by reference.

[0024] The polyamines suitable in the practice of the invention includethose having the formula:

R₁—N¹(R₂)—(CH₂)x-N²H—Q—N³H—(CH₂)y-N⁴(R₃)—R₄  (I)

[0025] wherein: R₁, R₂, R₃ and R₄ may be the same or different and areH, alkyl, cycloalkyl or aralkyl having from 1 to 12 carbon atoms, or aheterocyclic group having from 3 to 10 atoms wherein the hetero atom issaid N¹ or N⁴;

[0026] Q is a cycloalkyl group having from 3 to 10 carbon atoms;

[0027] x is an integer from 3 to 6, inclusive;

[0028] and y is an integer from 3 to 6, inclusive;

[0029] or (II) a salt thereof with a pharmaceutically acceptable acid.

DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention is predicated on the discovery thatpolyamines of the above formula act to inhibit the potential for thelarge and small intestines to contract. While not wishing to be bound byany theory as to the mechanism of action of the polyamines as inhibitorsof this action of the intestines, it is hypothesized that the polyaminesfunction via a receptor-dependent regulation mechanism whereby themyoelectric activity of the muscle tissue of the colon and smallintestine and the secretion of fluid and electrolytes by these organsare modulated. In addition, some of these above effects may be directlyor indirectly mediated through the release of nitric oxide or throughthe activation of nitric oxide synthase.

[0031] For each of the utilities mentioned herein, the amount requiredof active agent, the frequency and mode of its administration will varywith the identity of the agent concerned and with the nature andseverity of the condition being treated and is, of course, ultimately atthe discretion of the responsible physician or veterinarian. In general,however, a suitable dose of agent will lie in the range of about 0.0001mg to about 500 mg per kilogram of mammal body weight being treated.Administration by the parenteral route (intravenously, intradermally,intraperitoneally, intramuscularly or subcutaneously) is preferred for aperiod of time of from one to ten days; although the agent may also beadministered orally. For chronic problems, the drug is administered asneeded, subcutaneously, intravenously, or orally.

[0032] While it is possible for the agents to be administered as the rawsubstances, it is preferable, in view of their potency, to present themas a pharmaceutical formulation. The formulations of the presentinvention, both for veterinary and human use, comprise the agenttogether with one or more acceptable carriers therefor and optionallyother therapeutic ingredients. The carrier(s) must be “acceptable” inthe sense of being compatible with the other ingredients of theformulation and not deleterious to the recipient thereof. Desirably, theformulations should not include oxidizing agents and other substanceswith which the agents are known to be incompatible. The formulations mayconveniently be presented in unit dosage form and may be prepared by anyof the methods well known in the art of pharmacy. All methods includethe step of bringing into association the agent with the carrier whichconstitutes one or more accessory ingredients. In general, theformulations are prepared by uniformly and intimately bringing intoassociation the agent with the carrier(s) and then, if necessary,dividing the product into unit dosages thereof.

[0033] Formulations suitable for parenteral administration convenientlycomprise sterile aqueous preparations of the agents which are preferablyisotonic with the blood of the recipient. Suitable such carriersolutions include phosphate buffered saline, saline, water, lactatedringers or dextrose (5% in water). Such formulations may be convenientlyprepared by admixing the agent with water to produce a solution orsuspension which is filled into a sterile container and sealed againstbacterial contamination. Preferably, sterile materials are used underaseptic manufacturing conditions to avoid the need for terminalsterilization.

[0034] The pharmaceutical compositions according to the invention arethose which are suitable for enteral, such as oral, administration andfor parenteral, such as subcutaneous, administration to warm-bloodedanimals, especially humans, and which contain the pharmacologicallyactive substance on its own or together with a pharmaceuticallyacceptable carrier. The dosage of the active substance depends on thespecies of warm-blooded animal and on the age and individual conditionthe illness to be treated and also on the mode of administration.

[0035] The novel pharmaceutical preparations contain from approximately10% to approximately 95%, and preferably from approximately 20% toapproximately 90%, of the active substance. Pharmaceutical compositionsaccording to the invention can, for example, be in unit dose form, suchas drages, tablets, capsules, suppositories or ampoules, and containfrom approximately 0.05 g to approximately 10.0 g, and preferably fromapproximately 0.3 g to approximately 1.0 g, of the active ingredient.

[0036] The pharmaceutical compositions of the present invention aremanufactured in a manner known per se, for example, by means ofconventional mixing, granulating, confectioning, dissolving orlyophilizing processes. Pharmaceutical compositions for oral use can beobtained by combining the active substance with one or more solidcarriers, if desired, granulating a resulting mixture and processing themixture or granulate, if desired or necessary after the addition ofsuitable adjuncts, to form tablets or dragee cores. In so doing, theycan also be incorporated into plastic carriers which release the activesubstances or allow them to diffuse in controlled amounts.

[0037] Suitable carriers are especially fillers such as guars, forexample, lactose, saccharose, mannitol or sorbitol, cellulosepreparations and/or calcium phosphates, for example, tricalciumphosphate or calcium hydrogen phosphate, also potato starch, gelatin,tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodiumcarboxymethylcellulose and/or polyvinylpyrrolidone, and/or, if desired,disintegrators such as the above-mentioned starches, also carboxymethylstarch, cross-linked polyvinylpyrrolidone, agar, alginic acid or a saltthereof such as sodium alginate. Adjuncts are especially useful, such asflow-regulating and lubricating agents, for example, silica, talc,stearic acid or salts thereof such as magnesium or calcium stearate,and/or polyethylene glycol. Dragee cores are provided with suitablecoatings that are, if desired, resistant to gastric juice, there beingused, inter alia, concentrated sugar solutions which optionally containgum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/ortitanium dioxide, lacquer solutions in suitable organic solvents orsolvent mixtures or, for the manufacture of coatings that are resistantto gastric juice, solutions of suitable cellulose, preparations such asacetylcellulose phthalate or hydroxypropylmethylcellulose phthalate.Coloring substances or pigments can be added to the tablets or coatings,for example for the purpose of identification or for indicatingdifferent doses of active substance.

[0038] Other orally administrable pharmaceutical compositions aredry-filled capsules made of gelatin and also soft, sealed capsules madeof gelatin and a plasticizer such as glycerol or sorbitol. Thedry-filled capsules may contain the active ingredient in the form of agranulate, for example, in admixture with fillers such as corn starch,binders and/or glidants such as talc or magnesium stearate andoptionally, stabilizers. In soft capsules, the active ingredient ispreferably dissolved or suspended in suitable liquids or wax-likesubstances such as fatty oils, paraffin oil or polyethylene glycols, itbeing possible also for stabilizers to be added. Other forms of oraladministration are, for example, syrups prepared in a customary mannerthat contain the active ingredient in, for example, suspended form andin a concentration of approximately from 5% to 20%, and preferablyapproximately 10%, or in a similar concentration that provides asuitable single dose when administered, for example, in measures of 5 or10 ml. Also suitable are, for example, powdered or liquid concentratesfor preparing shakes, for example, in milk. Such concentrates can alsobe packed in single-dose quantities.

[0039] Particularly suitable dosage forms for parenteral administrationare sterile aqueous solutions of an active ingredient in water-solubleform, for example, a water-soluble salt, or sterile aqueous injectionsuspensions which contain substances increasing the viscosity, forexample, sodium, carboxymethyl cellulose, sorbitol and/or dextran, andoptionally stabilizers. In addition, the active ingredient, with orwithout adjuvants, can also be in lyophilized form and brought intosolution prior to parenteral administration by the addition of suitablesolvents.

[0040] Such formulations may optionally contain one or more additionalingredients among which may be mentioned preservatives, such as methylhydroxybenzoate, chlorocresol, metacresol, phenol and benzalkoniumchloride. Such materials are of special value when the formulations arepresented in multi-dose containers.

[0041] Buffers may also be included to provide a suitable pH value forthe formulation and suitable materials include sodium phosphate andacetate. Sodium chloride or glycerin may be used to render a formulationisotonic with the blood. If desired, the formulation may be filled intothe containers under an inert atmosphere such as nitrogen or may containan anti-oxidant and are conveniently presented in unit dose ormulti-dose form, for example, in a sealed ampoule.

[0042] It will be appreciated that while the agents described hereinform acid addition salts and carboxyl acid salts, the biologicalactivity thereof will reside in the agent itself. These salts may beused in human and in veterinary medicine and presented as pharmaceuticalformulations in the manner and in the amounts (calculated as the base)described herein above, and it is then preferable that the acid moietybe pharmacologically and pharmaceutically acceptable to the recipient.Examples of such suitable acids include (a) mineral acids: hydrochloric,hydrobromic, phosphoric, metaphosphoric, and sulfuric acids; (b) organicacids: tartaric, acetic, citric, malic, lactic, fumaric, benzoic,glycollic, gluconic, gulonic, succinic and arylsulfonic, for example,p-toluenesulfonic acids.

[0043] In compounds of the invention, R1 and R2 are preferably methyl,ethyl, propyl, benzyl, etc., it being understood that the term “aralkyl”is intended to embrace any aromatic group, the chemical and physicalproperties of which do not adversely affect the efficacy and safety ofthe compound for therapeutic applications. Preferred, however, are thehydrocarbyl aralkyl groups, i.e., comprised only of C and H atoms.

[0044] Polyamines of the above formula are synthesized according to themethods described in application Ser. No. 07/210,520 filed Jun. 23,1988, now U.S. Pat. No. 5,091,576, and Ser. No. 07/870,441 filed Oct. 9,1991, the entire contents and disclosures of both of which areincorporated herein by reference.

[0045] The invention is illustrated by the following non-limitingexamples.

EXAMPLE 1 Synthesis of CHX(3,4,3)-trans

[0046] Referring to FIG. 1,N,N′-Bis[3-(ethylamino)propyl]-trans-1,4-cyclohexanediaminetetrahydrochloride [CHX(3,4,3)-trans, 1] was assembled via a fragmentsynthesis (Scheme 1) Disulfonamide 3, available in 90% yield fromreaction of trans- 1,4-diaminocyclohexane (2) with mesitylenesulfonylchloride (2 equiv) under biphasic conditions, was alkylated withN-(3bromopropyl)-N-ethylmesitylenesulfonamide (2 equiv, NaH, DMF) toprovide protected polyamine 4 in 73% yield. The amino groups of 4 wereunmasked with 30% HBr in acetic acid and phenol, furnishing the finalproduct 1 in 64% yield.

EXAMPLE 2 Castor Oil-induced Diarrhea in Rats

[0047] Male Sprague-Dawley rats (350-400 g, Harlan Sprague-Dawley,Indianapolis, Ind.) were fasted overnight in hanging wire cages andallowed free access to water. A typical experiment involved 20 rats: 5untreated controls and 5 pretreated with polyamine analogues at each ofthree doses (typically equivalent on a molar basis to 1, 5, 10, or 25mg/kg of DEHSPM) as either a sc injection or a po gavage 30 min prior tocastor oil. All animals were then challenged with castor oil (purchasedfrom a local drugstore) as a gastric gavage of 5 mL/kg of body weight att=0 and monitored for the onset and duration of diarrhea at 30-minintervals for a 6-h period during which they received no food or water(Eaker, E. Y.; Bixler, G. B.; Mathias, J. R., J. Pharm Exp. Ther. 1988,246, 786-789). Onset of diarrhea for the control rats was between 30 and90 min and lasted for at least 6 h. The animal weight and stool weightwere recorded at 2, 4, and 6 h.

EXAMPLE 3 Irritable Bowel Syndrome in Rats

[0048] Male Sprague-Dawley rats (200-350 g, Harlan Sprague-Dawley,Indianapolis, Ind.) were housed in hanging wire cages in a temperature-and humidity-controlled room with a 12hour light/dark cycle; the animalswere fasted overnight (unless non-fasted rats were utilized inexperiments involving PO administration) and allowed free access towater. Animal care and experimental procedures were approved by theInstitutional Animal Care and Use Committee. A typical experimentinvolved 20 rats: 5 untreated controls and 5 pretreated with polyamineanalogues at each of three doses as either a SC injection or a PO gavage30 min prior to commencement of the stress. All animals were then housedin individual polycarbonate cages containing a clear 70×50 mm Pyrexcrystallization dish inverted in the center. To begin the stress, waterwas added to each cage to within 0.5 cm of the top of the Pyrex dish.The fecal output of the animals was recorded at 30-min intervals for a6-h period, during which they received no food or water. Stool outputwas expressed as the number of fecal pellets excreted over the 6-hcollection period. Percent reduction was calculated by dividing the meanvalue from the treated animals (T) by the mean value from the controlanimals (C), subtracting the resulting quotient from 1.0, andmultiplying by 100 [i.e., (1.0−T/C)×100].

EXAMPLE 4 Efficacy of CHX(3,4,3)-trans in Rats

[0049] CHX(3,4,3)-trans (1) was effective in the castor oil-induceddiarrhea model. When administered sc at doses ranging from 0.0078 mg/kgto 24.9 mg/kg, there was a significant reduction in weight loss at alldose levels (P<0.005 to P<0.001, Table 1). In addition, the compoundalso significantly decreased stool output relative to controls at scdoses of 0.0156 mg/kg to 24.9 mg/kg (P<0.001 for all doses, Table 1).

[0050] CHX(3,4,3)-trans was also remarkably effective at controllingIBS. When administered sc at doses ranging from 0.03 mg/kg to 0.99mg/kg, there was a significant reduction in fecal output at all doselevels (P<0.001, Table 2). Oral administration of this compound alsodiminished fecal output at doses≧5 mg/kg (P≦0.001, Table 2). TABLE 1Antidiarrheal Activity of Polyamine Analogues Predicated on a (3,4,3)Backbone^(a) compd. structure/ dose 15 per kg weight P- % stool P- % no,abbreviation mg :mol n loss value^(c) reduction^(d) output^(e) value^(c)reduction^(d) 1 0 0 20 11.6 ∀ 3.7 — — 4.9 2.0 — — 0.0078 0.0181  5  8.8∀ 1.2 <0.005 24 3.6 ∀ 1.3 >0.05  NS 0.0156 0.036  5  3.1 ∀ 1.2 <0.001 730.4 ∀ 0.9 <0.001  92 0.03125 0.0726  5  4.2 ∀ 1.7 <0.001 64 0 ∀ 0 <0.001100 0.625 0.145  5  3.7 ∀ 0.8 <0.001 68 0 ∀ 0 <0.001 100 0.125 0.29  5 4.1 ∀ 0.7 <0.001 65 0 ∀ 0 <0.001 100 0.25 0.58  4  2.7 ∀ 0.7 <0.001 770 ∀ 0 <0.001 100 0.5 1.16  5  2.9 ∀ 0.1 <0.001 75 0 ∀ 0 <0.001 100 0.992.3  9  2.7 ∀ 1.3 <0.001 77 0 ∀ 0 <0.001 100 4.97 11.6  5  1.6 ∀ 1.1<0.001 86 0 ∀ 0 <0.001 100 24.87 57.8  5  0.8 ∀ 0.7 <0.001 93 0 ∀ 0<0.001 100

[0051] TABLE 2 Activity of Polyamine Analogues Against Stress-InducedIrritable Bowel Syndrome Dose, kg⁻¹ Stool P- % Structure/Abbreviation mgμmol n Output^(b) Value Reduction^(d)

0 0.03125 0.0625 0.125 0.25 0.5 0.99 0 0.0726 0.145 0.29 0.58 1.16 2.310 5 5 5 5 5 5 5 # 11.3 ± 4.1 5.4 ± 2.1 2.2 ± 1.3 0.6 ± 0.9 0 ± 0 0 ± 00 ± 0 — 0.001 <0.001 <0.001 <0.001 <0.001 <0.001 —52.2 80.5 94.7 100 100100

What is claimed is:
 1. An anti-diarrheal or gastrointestinalanti-spasmodic pharmaceutical composition comprising [A] an effectiveamount of a compound having the formula:R₁—N¹(R₂)—(CH₂)x-N²H—Q—N³H—(CH₂)y-N⁴(R₃)—R₄  (I) wherein: R₁, R₂, R₃ andR₄ may be the same or different and are H, alkyl, cycloalkyl or aralkylhaving from 1 to 12 carbon atoms, or a heterocyclic group having from 3to 10 atoms wherein the hetero atom is said N¹ or N⁴; Q is a cycloalkylgroup having from 3 to 10 carbon atoms; x is an integer from 3 to 6,inclusive; and y is an integer from 3 to 6, inclusive; or (II) a saltthereof with a pharmaceutically acceptable acid; and [B] apharmaceutically acceptable carrier therefor.
 2. A composition accordingto claim 1 wherein Q is connected either cis or trans as the (1,2),(1,3), (1,4), (1,5) or (1,6) isomer.
 3. A composition according to claim1 wherein Q is cyclohexyl.
 4. A composition according to claim 1 whereinx is 3 and y is
 3. 5. A composition according to claim 1 wherein x is 3,y is 3, R₁ and R₃ are both H and R₂ and R₄ are both ethyl.
 6. Acomposition according to claim 1 wherein Q is cyclohexyl; x and y are 3;R₁ and R₃ are both H, and R₂ and R₄ are both ethyl.
 7. A compositionaccording to claim 6 wherein said polyamine is the trans (1,4) isomer.8. A method of treating diarrhea or gastrointestinal spasm in a human ornon-human animal in need thereof comprising administering to said animalan effective amount of a compound having the formula:R₁—N¹(R₂)—(CH₂)x-N²H—Q—N³H—(CH₂)y-N⁴(R₃)—R₄  (I) wherein: R₁, R₂, R₃ andR₄ may be the same or different and are H, alkyl, cycloalkyl or aralkylhaving from 1 to 12 carbon atoms, or a heterocyclic group having from 3to 10 atoms wherein the hetero atom is said N¹ or N⁴; Q is a cycloalkylgroup having from 3 to 10 carbon atoms; x is an integer from 3 to 6,inclusive; and y is an integer from 3 to 6, inclusive; or (II) a saltthereof with a pharmaceutically acceptable acid.
 9. A method accordingto claim 8 wherein Q is connected either cis or trans as the (1,2),(1,3), (1,4), (1,5) or (1,6) isomer.
 10. A method according to claim 8wherein Q is cyclohexyl.
 11. A method according to claim 8 wherein x is3 and y is
 3. 12. A method according to claim 8 wherein x is 3, y is 3,R₁ and R₃ are both H and R₂ and R₄ are both ethyl.
 13. A methodaccording to claim 8 wherein Q is cyclohexyl; x and y are 3; R₁ and R₃are both H, and R₂ and R₄ are both ethyl.
 14. A method according toclaim 13 wherein said polyamine is the trans (1,4) isomer.